Effects of the phytoestrogen genistein on the development of the reproductive system of Sprague Dawley rats

OBJECTIVES: Genistein is known to influence reproductive system development through its binding affinity for estrogen receptors. The present study aimed to further explore the effect of Genistein on the development of the reproductive system of experimental rats. METHODS: Eighteen post-weaning female Sprague Dawley rats were divided into the following groups: (i) a control group that received vehicle (distilled water and Tween 80); (ii) a group treated with 10 mg/kg body weight (BW) of Genistein (Gen 10); and (iii) a group treated with a higher dose of Genistein (Gen 100). The rats were treated daily for three weeks from postnatal day 22 (P22) to P42. After the animals were sacrificed, blood samples were collected, and the uteri and ovaries were harvested and subjected to light microscopy and immunohistochemical study. RESULTS: A reduction of the mean weekly BW gain and organ weights (uteri and ovaries) were observed in the Gen 10 group compared to the control group; these findings were reversed in the Gen 100 group. Follicle stimulating hormone and estrogen levels were increased in the Gen 10 group and reduced in the Gen 100 group. Luteinizing hormone was reduced in both groups of Genistein-treated animals, and there was a significant difference between the Gen 10 and control groups (p<0.05). These findings were consistent with increased atretic follicular count, a decreased number of corpus luteum and down-regulation of estrogen receptors-a in the uterine tissues of the Genistein-treated animals compared to the control animals. CONCLUSION: Post-weaning exposure to Genistein could affect the development of the reproductive system of ovarian-intact experimental rats because of its action on the hypothalamic-pituitary-gonadal axis by regulating hormones and estrogen receptors.

Expression of TGF-β1 in the blood during fracture repair in an estrogen-deficient rat model

OBJECTIVES: Previous studies have reported that osteoporosis due to estrogen deficiency influences fracture healing. Transforming growth factor (TGF-b) has been found to be involved in fracture healing via the regulation of the differentiation and activation of osteoblasts and osteoclasts. The current study aimed to determine the effects of estrogen on the expression of TGF-β1 during fracture healing in ovariectomized rats. METHODS: Thirty female Sprague-Dawley rats weighing 200-250 g were assigned to: (i) a sham-operated group that was given a normal saline; (ii) an ovariectomized control group that was given a normal saline; or (iii) an ovariectomized + estrogen (100 mg/kg/day) group that was treated with conjugated equine estrogen. The right femur of all rats was fractured, and a Kirschner wire was inserted six weeks post-ovariectomy. Treatment with estrogen was given for another six weeks post-fracture. At the end of the study, blood samples were taken, and the right femur was harvested and subjected to biomechanical strength testing. RESULTS: The percentage change in the plasma TGF-β1 level before treatment was significantly lower in the ovariectomized control and estrogen groups when compared with the sham group (p<0.001). After six weeks of treatment, the percentage change in the plasma TGF-β1 level in the estrogen group was significantly higher compared with the level in the ovariectomized control group (p = 0.001). The mean ultimate force was significantly increased in the ovariectomized rats treated with estrogen when compared with the ovariectomized control group (p = 0.02). CONCLUSION: These data suggest that treatment with conjugated equine estrogen enhanced the strength of the healed bone in estrogen-deficient rats by most likely inducing the expression of TGF-β1.

Piper sarmentosum enhances fracture healing in ovariectomized osteoporotic rats: a radiological study

INTRODUCTION: Osteoporotic fractures are common during osteoporotic states. Piper sarmentosum extract is known to possess antioxidant and anti-inflammatory properties. OBJECTIVES: To observe the radiological changes in fracture calluses following administration of a Piper sarmentosum extract during an estrogen-deficient state. METHODS: A total of 24 female Sprague-Dawley rats (200-250 g) were randomly divided into 4 groups: (i) the sham-operated group; (ii) the ovariectomized-control group; (iii) the ovariectomized + estrogen-replacement therapy (ovariectomized-control + estrogen replacement therapy) group, which was supplemented with estrogen (100 μg/kg/day); and (iv) the ovariectomized + Piper sarmentosum (ovariectomized + Piper sarmentosum) group, which was supplemented with a water-based Piper sarmentosum extract (125 mg/kg). Six weeks after an ovariectomy, the right femora were fractured at the mid-diaphysis, and a K-wire was inserted. Each group of rats received their respective treatment for 6 weeks. Following sacrifice, the right femora were subjected to radiological assessment. RESULTS: The mean axial callus volume was significantly higher in the ovariectomized-control group (68.2 + 11.74 mm³) than in the sham-operated, estrogen-replacement-therapy and Piper sarmentosum groups (20.4 + 4.05, 22.4 + 4.14 and 17.5 + 3.68 mm³, respectively). The median callus scores for the sham-operated, estrogen-replacement-therapy and Piper sarmentosum groups had median (range, minimum - maximum value) as 1.0 (0 - 2), 1.0 (1 - 2) and 1.0 (1 - 2), respectively, which were significantly lower than the ovariectomized-control group score of 2.0 (2 - 3). The median fracture scores for the sham-operated, estrogen-replacement-therapy and Piper sarmentosum groups were 3.0 (3 - 4), 3.0 (2 - 3) and 3.0 (2 - 3), respectively, which were significantly higher than the ovariectomized-control group score of 2.0 (1 - 2) (p<0.05). CONCLUSION: The Piper sarmentosum extract improved fracture healing, as assessed by the reduced callus volumes and reduced callus scores. This extract is beneficial for fractures in osteoporotic states.

The effect of eurycoma longifolia jack on spermatogenesis in estrogen-treated rats

Introduction: There is little data concerning the ability of Eurycoma longifolia Jack (EL) to reverse the inhibitory effects of estrogen on testosterone production and spermatogenesis. The aim of the present study was to determine the effect of EL on testicular histology and sperm count in estrogen-treated male rats. Methods: Adult male Sprague-Dawley rats weighing 200-250 g were divided into four groups of six rats each. Group A (control) was given solvent in the same manner as the treated groups were given EL. Group B was treated with EL (8 mg/kg body weight) orally. Group C was treated with estradiol (E2) (intramuscular dose of 500 ìg/kg body weight), and group D received a combined treatment of oral EL and intramuscular E2. After fourteen consecutive days of treatment, rats from all groups were sacrificed and subjected to spermatogenic and epididymal sperm cell counts. Results: The spermatogenic cell count in the E2-treated group was significantly decreased as compared to the control (p < 0.05) and EL+E2-treated groups (p < 0.05). A similar finding was found for the epididymal sperm count; the E2-treated group had a significant decrease in the count compared to the control (p < 0.05) and EL+E2-treated groups (p < 0.05). Rats that were treated with EL alone exhibited significantly higher sperm counts and sperm motility when compared to the control group (p < 0.05). Conclusions: EL extract acts as a potential agent for reversing the effects of estrogen by increasing spermatogenesis and sperm counts in rats after fourteen consecutive days of treatment.

Fresh soy oil protects against vascular changes in an estrogen-deficient rat model: an electron microscopy study

OBJECTIVE: To observe the effects of consuming repeatedly heated soy oil on the aortic tissues of estrogen-deficient rats. METHODS: Thirty female Sprague Dawley rats (200- 250 g) were divided equally into five groups. One group served as the normal control (NC) group. The four treated groups were ovariectomized and were fed as follows: 2 percent cholesterol diet (OVXC); 2 percent cholesterol diet + fresh soy oil (FSO); 2 percent cholesterol diet + once-heated soy oil (1HSO); and 2 percent cholesterol diet + five-times-heated soy oil (5HSO). After four months, the rats were sacrificed, and the aortic tissues were obtained for histological studies. RESULTS: After four months of feeding, the NC, FSO and 1HSO groups had a lower body weight gain compared to the OVXC and 5HSO groups. The tunica intima/media ratio in the 5HSO group was significantly thicker (p < 0.05) compared to the NC, OVXC and FSO groups. Electron microscopy showed that endothelial cells were normally shaped in the FSO and NC groups but irregular in the 1HSO and 5HSO groups. A greater number of collagen fibers and vacuoles were observed in the 5HSO group compared to the other treatment groups. CONCLUSIONS: Fresh soy oil offered protection in the estrogen-deficient state, as these rats had similar features to those of the NC group. The damage to the tunica intima and the increase in the ratio of tunica intima/media thickness showed the deleterious effect of consuming repeatedly heated soy oil in castrated female rats.

Development and evaluation of 'neck pain and functional limitation scale': A validation study in the Asian context.

Background :There is a paucity of literature on validated outcome measurement tools for evaluation of neck pain and related disability in the Asian context. Aim :The main aim of the present study was to design a new tool called neck pain functional limitation scale (NPFLS) for measuring disability related to neck pain and observe its reliability, concurrent validity and criterion validity. Setting and design :This study was performed at the institutional hospital. Materials and methods :A total of 157 subjects (neck pain group) and 25 control subjects (control group) without neck pain were recruited for this study. NPFLS was framed as a new tool for this study, which consisted of 5 domains - pain intensity, activities of daily living, social activities, functional activities and psychological factors. Neck Bournemouth questionnaire (NBQ) was used as a gold standard to measure the concurrent validity and criterion validity of the NPFLS. Statistical analysis :Criterion validity and concurrent validity between the neck Bournemouth questionnaire (NBQ) and NPFLS scores were tested statistically using Mann-Whitney U test and Spearman correlation test. The reliability was tested by examining the internal consistency to calculate the Cronbach's alpha value for each item in NPFLS. Results : No significant difference between NPFLS and NBQ was observed using Mann-Whitney U Test, with P value greater than 0.05 (P= 0.557). Besides that, NPFLS had a high concurrent validity (r= 0.916) and good internal consistency with high Cronbach's alpha value of (r= 0.948), which demonstrated strong correlation between the items of NPFLS and NBQ. Conclusion : NPFLS demonstrated good reliability, high concurrent validity and criterion validity in this study. NPFLS can be used to assess neck pain and disability among patients with neck pain.

The effects of palm oil tocotrienol-rich fraction supplementation on biochemical parameters, oxidative stress and the vascular wall of streptozotocin-induced diabetic rats

OBJECTIVE: This study examined the effects of palm oil tocotrienol-rich fractions on streptozotocin-induced diabetic rats. METHODS: Animals were divided into three groups: (i) normal non-diabetic (NDM), (ii) diabetic treated (tocotrienol-rich fractions - TRF) and (iii) diabetic untreated (non-TRF). The treatment group received oral administration of tocotrienol-rich fractions (200 mg/kg body weight) daily for eight weeks. The normal non-diabetic and the diabetic untreated groups were fed standard rat feed. Blood glucose and lipid profiles, oxidative stress markers and morphological changes of the thoracic aorta were evaluated. RESULTS: Tocotrienol-rich fractions treatment reduced serum glucose and glycated hemoglobin concentrations. The tocotrienol-rich fractions group also showed significantly lower levels of plasma total cholesterol, low-density lipoprotein cholesterol, and triglyceride, as compared to the untreated group. The tocotrienol-rich fractions group had higher levels of high-density lipoprotein cholesterol, as compared to the untreated group. Superoxide dismutase activity and levels of vitamin C in plasma were increased in tocotrienol-rich fractions-treated rats. The levels of plasma and aorta malondealdehyde + 4-hydroxynonenal (MDA + 4-HNE) and oxidative DNA damage were significant following tocotrienol-rich fractions treatment. Electron microscopic examination showed that the normal morphology of the thoracic aorta was disrupted in STZ-diabetic rats. Tocotrienol-rich fractions supplementation resulted in a protective effect on the vessel wall. CONCLUSION: These results show that tocotrienol-rich fractions lowers the blood glucose level and improves dyslipidemia. Levels of oxidative stress markers were also reduced by administration of tocotrienol-rich fractions. Vessel wall integrity was maintained due to the positive effects mediated by tocotrienol-rich fractions.

Multiple vascular anomalies in the abdomen: a gross anatomical study / Múltiples anomalías vasculares en el abdomen: un estudio anatómico macroscópico

Knowledge of the branching pattern of the abdominal aorta is clinically important for any abdominal surgeon operating on parts of the gut or neighboring structures like the suprarenals, spleen, pancreas, liver, kidneys and ureter. The presence of abnormal inferior phrenic artery associated with aberrant branch from the celiac trunk supplying the pancreas and duodenum is a rare anomaly. In the present case, we observed four branches of the celiac artery i.e. (a) left gastric artery (b) common hepatic artery (c) splenic artery and (d) an aberrant branch, which took a course inferiorly towards the pancreas. The aberrant artery supplied the body of the pancreas and gave a branch which supplied the horizontal part of the duodenum and then entered the transverse mesocolon to supply the hepatic flexure and some portions of the ascending and the transverse colon. The inferior phrenic artery was absent on the left side. Concomitant anomalies of such type are to be kept in mind by the surgeon, while operating cases of carcinoma head of pancreas and performing kidney transplantations.

El conocimiento del patrón de ramificación de la aorta abdominal es clínicamente importante para cualquier cirujano abdominal que opere en partes del intestino o estructuras vecinas, como glándulas suprarenales, bazo, páncreas, hígado, riñones y uréteres. La presencia anormal de la arteria frénica inferior asociada con una rama aberrante originada del tronco celiaco, supliendo el páncreas y duodeno, es una variación anatómica rara. En el presente caso, se observaron cuatro ramas de la arteria celiaca: (a) arteria gástrica izquierda (b) arteria hepática común (c) arteria esplénica y (d) una rama aberrante, que tuvo un curso inferior hacia el páncreas. La arteria aberrante suministraba irrigación al cuerpo del páncreas y daba una rama para la parte horizontal del duodeno para luego entrar en el mesocolon transverso para irrigar la flexura hepática y algunas partes del colon ascendente y transverso. La arteria frénica inferior estaba ausente en el lado izquierdo. Anomalías concomitantes de este tipo deben ser consideradas por el cirujano, en casos de operación de carcinoma de cabeza de páncreas y la realización de trasplante renal.

Laser capture microscope: an innovative tool for single cell dissection.

Increasing strides have recently been made in the understanding of the biologic basis of health and disease, largely because of the current emphasis on investigations directed at a cellular level. This has necessitated the development and use of convenient, accurate and reliable research equipment. While it is not necessitated the need for every researcher or clinician to be fully knowledgeable of all the intricacies of the recent developments in the field of research equipment, basic awareness of new research tools is the critical to the understanding and analysis of results obtained from current studies. The laser capture microscope is one of the most recent research equipment that permits easy dissection and examination of single cells from tissues. This has enabled researchers to carry out specific studies on the behaviour of different cell populations in a tissue sample. This article aims to explain the basic principles of the usage of laser capture microscope for single cell dissection.

Ginger extract (Zingiber officinale) has anti-cancer and anti-inflammatory effects on ethionine-induced hepatoma rats

OBJECTIVE: To evaluate the effect of ginger extract on the expression of NFêB and TNF-á in liver cancer-induced rats. METHODS: Male Wistar rats were randomly divided into 5 groups based on diet: i) control (given normal rat chow), ii) olive oil, iii) ginger extract (100mg/kg body weight), iv) choline-deficient diet + 0.1 percent ethionine to induce liver cancer and v) choline-deficient diet + ginger extract (100mg/kg body weight). Tissue samples obtained at eight weeks were fixed with formalin and embedded in paraffin wax, followed by immunohistochemistry staining for NFêB and TNF-á. RESULTS: The expression of NFêB was detected in the choline-deficient diet group, with 88.3 ± 1.83 percent of samples showing positive staining, while in the choline-deficient diet supplemented with ginger group, the expression of NFêB was significantly reduced, to 32.35 ± 1.34 percent (p<0.05). In the choline-deficient diet group, 83.3 ± 4.52 percent of samples showed positive staining of TNF-á, which was significantly reduced to 7.94 ± 1.32 percent (p<0.05) when treated with ginger. There was a significant correlation demonstrated between NFêB and TNF-á in the choline-deficient diet group but not in the choline-deficient diet treated with ginger extract group. CONCLUSION: In conclusion, ginger extract significantly reduced the elevated expression of NFêB and TNF-á in rats with liver cancer. Ginger may act as an anti-cancer and anti-inflammatory agent by inactivating NFêB through the suppression of the pro-inflammatory TNF-á.

A study of the antioxidant effect of alpha lipoic acids on sperm quality

OBJECTIVE: Assisted reproductive techniques are useful in helping infertile couples achieve successful conception. Initial studies have shown that sperm cryopreservation, one step in assisted reproduction, causes a dramatic reduction in sperm quality. This has been attributed to, among other things, free radical activities. The aim of the present study was to minimize this oxidative attack by adding an antioxidant into the sperm microenvironment. Alpha lipoic acids were selected for this purpose for their efficient free radical scavenging properties and solubility in lipid and aqueous phases. METHODS: For this investigation, semen from six Boer bucks was pooled. Seminal analysis of the baseline prior to incubation of samples with different concentrations of Alpha lipoic acids (0.00625, 0.0125, 0.025, 0.05, 0.1 mmol/ml) was performed, and post-seminal analysis was conducted after a one-hour incubation. The comet assay was used to observe the effect of Alpha lipoic acids on sperm DNA integrity. Statistical analysis using an unpaired t-test with a significance level of p<0.05 was then performed. RESULTS: Our results indicate that the sperm motility rate was improved after incubation with Alpha lipoic acids at a concentration of 0.02 mmol/ml. This concentration was also capable of reducing DNA damage. CONCLUSION: In conclusion, Alpha lipoic acids renders cryoprotection to sperm, thereby improving sperm quality.

Ossification of caroticoclinoid ligament and its clinical importance in skull-based surgery

CONTEXT: The medial end of the posterior border of the sphenoid bone presents the anterior clinoid process (ACP), which is usually accessed for operations involving the clinoid space and the cavernous sinus. The ACP is often connected to the middle clinoid process (MCP) by a ligament known as the caroticoclinoid ligament (CCL), which may be ossified, forming the caroticoclinoid foramen (CCF). Variations in the ACP other than ossification are rare. The ossified CCL may have compressive effects on the internal carotid artery. Thus, anatomical and radiological knowledge of the ACP and the clinoid space is also important when operating on the internal carotid artery. Excision of the ACP may be required for many skull-based surgical procedures, and the presence of any anomalies such as ossified CCL may pose a problem for neurosurgeons. CASE REPORT: We observed the presence of ossified CCL in a skull bone. A detailed radiological study of the CCL and the CCF was conducted. Morphometric measurements were recorded and photographs were taken. The ACP was connected to the MCP and was converted into a CCF. Considering the fact that standard anatomy textbooks do not provide morphological descriptions and radiological evaluations of the CCL, the present study may be important for neurosurgeons operating in the region of the ACP.

CONTEXTO Y OBJETIVO: El extremo intermedio de la frontera posterior del hueso del sphenoid presenta el proceso anterior del clinoid (ACP), que está alcanzado generalmente para las operaciones que implican el espacio clinoidal y el sino cavernoso. El ACP es conectado a menudo con el proceso medio del clinoid (MCP) por un ligamento conocido como el ligamento del carotico-clinoid (CCL), que puede ser osificado, formando el agujero del clinoid del carotico (CCF). Las variaciones del ACP con excepción de la osificación, son raras. El CCL osificado puede tener efectos compresivos en la arteria carótida interna. El conocimiento así anatómico y radiológico del ACP y del espacio clinoidal es también importante mientras que funciona en la arteria carótida interna. La supresión del ACP se puede requerir para muchos cirugía basada cráneo y la presencia de cualquier anomalía tal como CCL osificado, puede plantear un problema para el neurocirujano. INFORME DEL CASO: Observamos la presencia de CCL osificado en un hueso del cráneo. Un estudio radiológico detallado del CCL y del CCF fue conducido. Las medidas morphometric fueron registradas y las fotografías fueron tomadas. El ACP fue conectado con el MCP y convertido en un CCF. En vista del hecho, los libros de textos de esa anatomía del estándar no proporcionan la descripción morfológica y la evaluación radiológica de CCL, el actual estudio puede ser importante para los neurocirujanos que funcionan en la región del ACP.

Anomalous posterior clinoid process and its clinical importance / Importancia clínica de anomalías en los procesos clinoides posteriores

The anterior, middle and the posterior clinoid processes are located in the middle cranial fossa of the skull. The posterior boundary of the pituitary fossa is formed by the dorsum sellae, the supero-lateral angles of which are raised to form the posterior clinoid process. Unlike the anterior clinoid process, the posterior clinoid process has received less attention in past research studies. The anatomy textbooks do not mention about the anomalies pertaining to the posterior clinoid process and the main source of information are the research reports. The present osteological study describes anomalous posterior clinoid process, detected in two human skulls, during routine undergraduate medical teaching and also discusses its clinical importance. The posterior clinoid process anomalies may be responsible for the altered arrangement of the tentorium cerebelli which is attached to it. The internal carotid artery is often explored by the neuro-surgeon and any bony abnormalities in its vicinity, would result in a difficult situation, while performing clinoidectomy operations. Anomalous posterior clinoid process, may compress the internal carotid artery. The close position of the superior petrosal sinus and the internal carotid artery to the posterior clinoid process, makes it vulnerable to injuries and thus it is important for the neuro-surgeons performing clinoidectomy operations. The anatomy of the posterior clinoid process may be important for neuro surgeons and radiologists in day to day clinical practice.

Los procesos clinoides anterior, medio y posterior se localizan en la fosa media del cráneo. El límite posterior de la fosa pituitaria o silla turca lo constituye la lámina cuadrilátera del esfenoides = dorsum sellae, o dorso de la silla turca, cuyos ángulos supero-laterales se levantan para formar el proceso clinoides posterior. Al contrario del proceso clinoides anterior, el proceso clinoides posterior ha recibido menos atención en los estudios del pasado. Los textos de anatomía no mencionan las anomalías del proceso clinoides posterior y las principales fuentes de conocimiento son los informes de investigaciones. El presente trabajo osteológico describe anomalías del proceso clinoides posterior que se encontraron en dos cráneos humanos, en el curso de enseñanza médica de pre-grado y también discute su valor e interés clínicos. Las anomalías del proceso clinoides posterior pueden ser responsables de alteraciones en la disposición del tentorium cerebelli al que se adhiere. Con frecuencia el neurocirujano explora la arteria carótida interna y las anomalías óseas en su vecindad pueden resultar en situaciones difíciles mientras se hacen clinoidectomías. Un proceso clinoides posterior anormal puede comprimir la arteria carótida interna. Las estrechas cercanías que hay entre el seno petroso superior y el proceso clinoides posterior con la arteria carótida interna, hacen que este vaso sea vulnerable a posibles lesiones; esto es fundamental para las clinoidectomías que ejecutan los neurocirujanos. Además, la anatomía del proceso clinoides posterior es de importancia no sólo para los neurocirujanos sino también para los radiólogos en su práctica clínica de todos los días.